CN101549008B - Application of cassia seed extract extracted continuously under high pressure in the preparation of blood lipid-lowering drugs - Google Patents
Application of cassia seed extract extracted continuously under high pressure in the preparation of blood lipid-lowering drugs Download PDFInfo
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Abstract
The present invention relates to application of high-pressure continuous extracted cassia seed extract in preparing hypolipemic medicine. The high-pressure continuance extracted cassia seed extract possess stronger hypolipemic pharmacological effects than extract by other extraction methods, and its effects in curing hyperlipemia is obviously better than the compound preparation-Wenstrandin, therefore having no need to match with other medicines to prepare cassia seed single preparation for hypolipemic application, and processing evident hypolipemic impression.
Description
Technical field
The present invention relates to the extract application in pharmaceutical field, the especially high pressure application of Semen Cassiae extract in the blood lipid-lowering medicine preparation of extraction continuously of Semen Cassiae.
Background technology
Hyperlipemia is a kind of common disease, is due to the human body lipid metabolism is lacked of proper care.Clinical Laboratory is serum cholesterol (TC), triglyceride (TG), (LDL) is too high for low density lipoprotein, LDL; High density lipoprotein (HDL) is tending towards descending or being lower than normal value.Hyperlipemia is easily brought out cardiovascular disease such as coronary heart disease, apoplexy, hypertension, also may cause diseases such as cholelithiasis, pancreatitis, senile dementia.At present, along with rapid economy development, great changes will take place for people's dietary structure, high heat food intake rates such as Animal fat increase, add the variation of the socio-psychological factors of rhythm of life due to accelerating, hyperlipidemia population increases, and the sickness rate of cardiovascular and cerebrovascular disease raises.Now, doctor trained in Western medicine adopts lipid lowerers such as simvastatin, atorvastatin, pravastatin, fluvastatin, lovastatin (main cholesterol reducing, slight triglyceride reducing) to treat more clinically.Though these medicines have effect for reducing blood fat, many side effect are also arranged, so medical circle is just making great efforts to develop the little blood lipid-lowering medicine of side effect.
Semen Cassiae is the dry mature seed of leguminous plant Semen Cassiae CassiaobtusifoliaL. or little Semen Cassiae C.toraL., is clinical conventional Chinese medicine, and Shennong's Herbal is classified it as top grade.Semen Cassiae is Semen Cassiae again, the Horse hoof Semen Cassiae, and false Semen phaseoli radiati, all there is cultivation in national most of area, and gathered behind the fruit maturation to autumn in main product Anhui, Guangxi, Zhejiang, Sichuan, its Herb is cut off dry, and gets seed.The surface of the seed yellow green, nature and flavor are sweet, bitter, salty, be slightly cold.Have liver heat removing and eyesight improving, the effect of loosening bowel to relieve constipation, modern study proof Semen Cassiae has blood pressure lowering, blood fat reducing, protects the liver and antibacterial isoreactivity, can be used as food again simultaneously, is the good raw material of health beverage.
In recent years, the Semen Cassiae compound preparation of existing clinically multiple treatment hyperlipemia, as (1) XUEZHINING WAN, the pill of making by Semen Cassiae, Fructus Crataegi, Folium Nelumbinis and Radix Polygoni Multiflori Preparata composition of prescription; (2) XUEZHILING PIAN: the tablet of making by Radix Polygoni Multiflori Preparata, Fructus Lycii, Rhizoma Polygonati, Fructus Crataegi, Semen Cassiae composition of prescription; (3) Colestid capsule: the capsule that forms by Fructus Crataegi (enucleation), Radix Polygoni Multiflori Preparata, Semen Cassiae, Folium Nelumbinis and Rhizoma Alismatis composition of prescription; (4) the glad recovering particles of fat: the granule that forms by Semen Cassiae, Folium Nelumbinis, Herb Gynostemmae Pentaphylli, Rhizoma Alismatis, Flos Lonicerae, Folium Ilicis and Folium Ginkgo composition of prescription; (5) defatting and poison expelling capsule: the capsule that forms by Radix Et Rhizoma Rhei, Semen Cassiae, Fructus Crataegi, Herba Artemisiae Scopariae, Rhizoma Alismatis, Fructus Gardeniae, Rhizoma Curcumae, Radix Polygoni Multiflori and Radix Bupleuri composition of prescription; (6) Fat-reducing are blood-circulation promoting capsule: the capsule that forms by Rhizoma Curcumae Longae, Herba cynodontis, Rhizoma Alismatis, Semen Cassiae and Radix Notoginseng composition of prescription; (7) Semen Cassiae capsule ' Pingzhi ': Semen Cassiae, Herba Artemisiae Scopariae, Radix Polygoni Multiflori, Herba Taxilli, vitamin C, vitamin B, nicotinic acid.Above-mentioned Semen Cassiae compound preparation is treated hyperlipemia effectively, has obtained satisfied effect, but these therapeutic effect are Semen Cassiaes and synergism is following produces for the generation of other drug compatibility, rather than the therapeutic effect that possessed of Semen Cassiae single medication.
At present, because the existing extracting method of pharmaceuticals industry still can not be complete with extracts active ingredients in the Semen Cassiae, limited the application in the treatment of hyperlipemia clinically of Semen Cassiae single preparations of ephedrine.
Summary of the invention
The purpose of this invention is to provide the application of Semen Cassiae extract in the blood lipid-lowering medicine preparation that high pressure extracts continuously.
Blood lipid-lowering medicine of the present invention is the medicine of cholesterol reducing, triglyceride, low density lipoprotein, LDL and high density lipoprotein increasing.
Blood lipid-lowering medicine of the present invention is the medicine of cholesterol reducing.
Blood lipid-lowering medicine of the present invention is the medicine of triglyceride reducing.
Blood lipid-lowering medicine of the present invention is the medicine that reduces low density lipoprotein, LDL.
Blood lipid-lowering medicine of the present invention is the medicine of high density lipoprotein increasing.
The extract of the Semen Cassiae that high pressure of the present invention extracts is continuously made by following method: depend on that the pine torch powder mixes immersion 30～60 minutes with ethanol by solid-liquid ratio 1: 10～30; Then the feed liquid of gained is sent in the high pressure extract device, feed liquid is passed through the annular working gap of high pressure extract device continuously under the effect of 30～90Mpa high pressure, on the collision ring of high speed impact in the high pressure extract device, make the cell wall rupture of Semen Cassiae, effective ingredient is spread in the solvent; Take out the feed liquid through HIGH PRESSURE TREATMENT at last, filter, remove the material slag, concentrating filter liquor, drying obtain the extract powder of Semen Cassiae extract.
The extracting method of the extract of the Semen Cassiae that the present invention can also extract continuously high pressure is done following the improvement: promptly Semen Cassiae circulates continuously under the effect of 30～90Mpa high pressure with the feed liquid of ethanol mixings gained and repeatedly passes through the annular working gap of high pressure extract device, so that the cell wall that makes Semen Cassiae helps further improving the Semen Cassiae extraction ratio of effective constituents through multiple impact.The persistent period of described continuous cyclic process is in the scope of 10～30min.The persistent period of best cyclic process continuously is in 20～30min scope.
The preferred solid-liquid ratio of Semen Cassiae and ethanol is in 1: 10～25 scope among the present invention, and best solid-liquid ratio is then in 1: 15～20 scope.
The preferable force value of high pressure extract of the present invention is in 40～90Mpa scope, and the optimum pressure value is in 50～80Mpa scope.
Concentration of ethanol described in the present invention is 50%～90%, and its preferable concentration is 60%～90%, and best concentration is 60%～80%.
Adopt high performance liquid chromatography-electrospray ionization mass spectrometry coupling method (HPLC-ESI-MS through the inventor
n) Semen Cassiae extract of the present invention is carried out the whole part analysis, from Semen Cassiae extract of the present invention, isolate following 10 kinds of effective ingredient: Semen Cassiae glycosides B
2(molecular weight 920), Semen Cassiae glycosides C
2(molecular weight 920), chrysophanol-tetrapyran glucoside (molecular weight 902), chrysophanol-1-O-β-D-glucopyranosyl-(1 → 3)-β-D-glucopyranosyl-(1 → 6)-β D-pyranglucoside (molecular weight 740), red sickle enzyme element-6-β-gentiobiose glycosides (molecular weight 596), red sickle enzyme element-1-O-gentiobiose glycosides (molecular weight 596), chrysophanol-1-O-β-gentiobiose glycosides (molecular weight 578); 1,2,6-trihydroxy-7,8-dimethoxy-3-tectoquinone (molecular weight 330); 2-hydroxyl-1,6,7,8-tetramethoxy-3-tectoquinone (molecular weight 358); 3-methyl-2,8-dihydroxy-1,6,7-trimethoxy anthraquinone (molecular weight 344).
The inventor adopts petroleum ether, ethyl acetate, n-butyl alcohol to carry out Semen Cassiae extract of the present invention to carry out branch extraction, get the extract at each chemical position of Semen Cassiae, carry out the research of blood fat reducing drug effect, find that n-butanol extract is the main blood lipid-lowering active fractions of Semen Cassiae.N-butanol extract to Semen Cassiae adopts high speed adverse current chromatogram (HSCCC) to carry out separation and purification, obtains 5 effective ingredient: 1,2,6, and 8-tetrahydroxy-7-methoxyl group-3-tectoquinone (molecular weight 316); 1,2,6-trihydroxy-7,8-dimethoxy-3-tectoquinone (molecular weight 330); 2-hydroxyl-1,6,7,8-tetramethoxy-3-tectoquinone (molecular weight 358); 2,6-dihydroxy-1,7,8-trimethoxy-3-tectoquinone (molecular weight 344); 1,2-dihydroxy-6,7,8-trimethoxy-3-tectoquinone (molecular weight 344).
Semen Cassiae extract of the present invention can be prepared into various dosage forms according to route of administration, comprises oral formulations and ejection preparation.Wherein oral formulations comprises: tablet, granule, capsule, oral liquid, pill etc.Available adjuvant can be conventional filleies such as starch, dextrin, cyclodextrin, sucrose, stearate when the preparation oral formulations.
Below by experiment and the result illustrate that the Semen Cassiae extract that adopts high pressure to extract continuously is applied to treat the effect of hyperlipemia as single preparations of ephedrine.
One, about the high pressure effective ingredient research of the Semen Cassiae extract of extraction continuously
(1) experiment material and experimental technique:
Exsiccant Semen Cassiae medical material (purchasing) in Guangzhou medical material company;
Instrument: LCQ Advantage liquid chromatograph/mass spectrometer (U.S. Thermo Finnigan company), high performance liquid chromatography (HPLC) pump (Japan, Jasco company), phenomenex ODS3 post (250mm * 4.6mm); Ion trap mass spectrometry, electrospray interface, work station and data handling system (U.S. Thermo Finnigan company).
Mass spectrum condition: electron spray voltage ± 5.0Kv; 200 ℃ of metal capillary temperature; Shell gas (N
2) flow: 60mL/min; Flow injection pump sample introduction, sample size 3 μ L/min; Scan mode: cation, anion scanning.
Chromatographic condition: Phenomenex ODS3 post (250mm * 4.6mm); Mobile phase: methanol-0.4% acetic acid solution gradient elution, elution time is 90min, and 0-5min changed to 35: 65 from 15: 85, and 5-65min changed to 50: 50 from 35: 65, and 65-80min changed to 100: 0 from 50: 50, continue to keep 10min at 100: 0 again; Volume flow 0.8mL/min; Detect wavelength: 254nm; Column temperature: 25 ℃.
(2) high pressure continuous extraction method
Take by weighing Semen Cassiae powder 50g, add 70% ethanol 1000ml and mix immersion 30～60 minutes, feed liquid is sent in the high pressure extract device then, feed liquid circulates continuously under the effect of 60Mpa high pressure and repeatedly passes through the annular working gap of high pressure extract device, so that make the cell wall process multiple impact of Semen Cassiae, described continuous circulation time remaining 30min, to take out through the feed liquid of HIGH PRESSURE TREATMENT, filter, reclaim ethanol, concentrate Semen Cassiae extractum.
(3) analysis of effective ingredient in the continuous Semen Cassiae extract that extracts of high pressure
Get high pressure and extract resulting extract employing high performance liquid chromatography-electrospray ionization mass spectrometry coupling method (HPLC-ESI-MS continuously
n) carrying out the whole part analysis, the HPLC chromatogram (Fig. 1) of gained and total cation flow graph (Fig. 2) carry out interpretation of mass spectra according to the charge-mass ratio (m/z) and the feature fragment of molecular ion peak, have confirmed 10 kinds of anthraquinone analog compounds (table 1) wherein.
The structural information and the relative retention time of 10 kinds of anthraquinone analog compounds of table 1
The extraction ratio of the total flavones of this method is 1.233%, and the extraction ratio of general anthraquinone is 0.089%, and the extraction ratio of total polysaccharides is 10.55%.
Two, the drug effect contrast experiment of the extract of different solvents
(1) experiment material
84 of male SD rats (quality certification SCXK (Guangdong) 2003-0002)
XUEZHIKANG JIAONANG (Beijing big dimension letter bio tech ltd, the accurate word of traditional Chinese medicines: Z10950029, lot number: 20080110)
Cholesterol reagent box (glad biotechnology research institute of Shanghai section, lot number: 20080301)
High density lipoprotein test kit (glad biotechnology research institute of Shanghai section, lot number: 20080301)
Low density lipoprotein, LDL test kit (glad biotechnology research institute of Shanghai section, lot number: 20080301)
Triglyceride test kit (glad biotechnology research institute of Shanghai section, lot number: 20080301)
High glucose and high fat feed formula: 81% normal feedstuff, 5% sodium cholate, 1.5% cholesterol, 2% yolk powder, 3% sugar, 10% Adeps Sus domestica, 25g mineral, 50g compound vitamin
Instrument: visible spectrophotometer (the general all purpose instrument company limited of analysing in Beijing)
(2) experimental technique
84 of male SD rats, adaptability are divided into 7 groups at random by body weight after raising a week, 12 every group, are respectively: normal control group, model control group, Xuezhikang matched group (0.11g/kgd
-1), the ligroin extraction group, the ethyl acetate extract group, the n-butanol extract group, the water extract group, feed in formal experiment first day and to carry out modeling to the high glucose and high fat feedstuff, and inject carbon tetrachloride vegetable oil mixed liquor (60% vegetable oil for each modeling group rat muscle, 40% carbon tetrachloride) 3ml/kg, each group gives the medicine filling stomach of corresponding dosage simultaneously, the normal control group gives the equal-volume normal saline, continuous 8 weeks, centrifugal blood (2000r/min, 10min) get serum, detect and respectively organize rat blood serum T-CHOL (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein, LDL (LDL) level, testing result sees Table 2:
Each extract of table 2 Semen Cassiae is to the influence of hyperlipidemia mice blood fat
Annotate:
#＜0.05,
##P＜0.01 is compared with normal group;
*P＜0.05,
*(corresponding diagram together) compared in P＜0.01 with model group
The result of table 2 shows: (1) model group with hyperlipemia and normal control group compare, and the TC (P＜0.01) that can significantly raise, TG (P＜0.05), LDL (P＜0.01) reduce HDL, and this method modeling success is described.(2) compare with the hyperlipemia group, n-butanol extract can significantly reduce TC (P＜0.01), TG (P＜0.05) and the LDL (P＜0.01) of hyperlipemia in mice, rising HDL, and its activity is strong than Xuezhikang (positive drug); Ligroin extraction can significantly reduce the TC (P＜0.05) of hyperlipemia in mice, and reduction rate is 14.97%; Water extract can significantly reduce the TG (P＜0.05) of hyperlipemia in mice, and reduction rate is 25.2%.All the other each extracts can reduce TC and TG to a certain extent, but compare with model group, and difference does not have significance meaning (P＞0.05).Therefore, the Semen Cassiae n-butanol extract is a blood lipid-lowering active fractions.
Three, the analysis of effective component in the n-butanol extract
(1) instrument and reagent
GS10A type high-speed counter-current chromatograph (politef post, internal diameter 1.6mm, column volume 230ml, rotating speed 0-1000r/min), the 8823A-UV Ultraviolet Detector, S-1007A pump, S-type monitor (New Technique Application Inst., Beijing City), the automatic fraction collector of BSZ-160 (Luxi, Shanghai instrument plant), automatically controlled tepidarium heater (20-100 ℃ of temperature control scope).
Waters 600 series of high efficiency chromatograph of liquid, Waters 2487 detectors, HS chromatographic data work station; Computer aided similarity evaluation system.
LCQ Advantage liquid chromatograph/mass spectrometer (U.S. Thermo Finnigan company), high performance liquid chromatography (HPLC) pump (Japan, Jasco company), Phenomenex ODS3 post (250mm * 4.6mm); Ion trap mass spectrometry, electrospray interface, work station and data handling system (U.S. Thermo Finnigan company).
Semen Cassiae (Guangzhou medical material company); Ethanol, acetic acid, ethyl acetate, normal hexane etc. are analytical pure; Methanol is chromatographically pure; Water is deionized water, WAHAHA pure water (mass spectrum is used).
(2) experimental technique
Separate solvent system: n-hexane/ethyl acetate/methanol=4: 1: 3: 2;
Separation chromatography condition: column volume 230ml, flow rate of mobile phase 1.5ml/min, rotating speed 750r/min, UV-detector wavelength 254nm, monitor sensitive volume 2A, range 200mV, chart speed 60cm/h.
(3) experimental result
After separating, HSCCC shows 6 main peaks (referring to Fig. 3), collect each component according to peak sequence with catcher, observing first big peak during according to collection is that mixture detects to find it is the mixture of the very approaching several chemical compounds of polarity through HPLC, because polarity very is difficult to separate near consequently going out the peak simultaneously.Component I, II, III, IV, V detect through HPLC and show and be one matter and purity greater than 95%.
Five other on adverse current chromatogram figure main peaks have been carried out the HPLC detection, and the purity of five components is respectively 95%, 98%, 96%, 95% and 96%.The HPLC of each peak gleanings analyzes shown in Fig. 4-1～4-5.These five peak compositions are repeatedly collected, and carried out discovering of mass spectral analysis and the cracked mechanism of mass spectrum respectively, these five compositions are respectively: 1,2,6, and 8-tetrahydroxy-7-methoxyl group-3-tectoquinone (molecular weight 316); 1,2,6-trihydroxy-7,8-dimethoxy-3-tectoquinone (molecular weight 330); 2-hydroxyl-1,6,7,8-tetramethoxy-3-tectoquinone (molecular weight 358); 2,6-dihydroxy-1,7,8-trimethoxy-3-tectoquinone (molecular weight 344); 1,2-dihydroxy-6,7,8-trimethoxy-3-tectoquinone (molecular weight 344).
Four, the drug effect contrast experiment of the extract of Different Extraction Method
(1) contrast experiment of the extraction ratio of Different Extraction Method
Exsiccant Semen Cassiae medical material is ground into powder, is divided into three groups, adopt following three kinds of extracting method to extract and measure the extraction ratio of total flavones, general anthraquinone and total polysaccharides respectively:
1. reflux extraction: take by weighing Semen Cassiae powder 50g, add 70% ethanol 1000ml, 70 ℃ are extracted 3.5h, filter, and reclaim ethanol, concentrate Semen Cassiae extractum; The extraction ratio of the total flavones of this method is 0.895%, and the extraction ratio of general anthraquinone is 0.071%, and the extraction ratio of total polysaccharides is 9.324%.
2. supersound extraction: take by weighing Semen Cassiae powder 50g, add 70% ethanol 1000ml, under the power of 500W, extract 40min, filter, reclaim ethanol, concentrate Semen Cassiae extractum; The extraction ratio of the total flavones of this method is 0.877%, and the extraction ratio of general anthraquinone is 0.069%, and the extraction ratio of total polysaccharides is 8.642%.
3. high pressure extracts continuously: take by weighing Semen Cassiae powder 50g, add 70% ethanol 1000ml and mix immersion 30～60 minutes, feed liquid is sent in the high pressure extract device then, feed liquid circulates continuously under the effect of 60Mpa high pressure and repeatedly passes through the annular working gap of high pressure extract device, so that make the cell wall process multiple impact of Semen Cassiae, described continuous circulation time remaining 30min, to take out through the feed liquid of HIGH PRESSURE TREATMENT, filter, reclaim ethanol, concentrate Semen Cassiae extractum.The extraction ratio of the total flavones of this method is 1.233%, and the extraction ratio of general anthraquinone is 0.089%, and the extraction ratio of total polysaccharides is 10.55%.
From The above results as seen, the extraction ratio of the total flavones of the Semen Cassiae extract that extracts continuously of high pressure, general anthraquinone, total polysaccharides is respectively 1.4 times, 1.3 times, 1.15 times of extract of reflux, extract.And reflux, extract, is close with the supersound extraction yield.And show from the HPLC finger printing shown in Fig. 5～7: the peak the HPLC finger printing of the Semen Cassiae extract that three kinds of extracting method obtain is similar, all concentrate on retention time 10-55min and 72-90min, this shows that the present invention does not change the effective ingredient in the Semen Cassiae.And in Fig. 3, show under the identical situation of Semen Cassiae content, each peak height is all than the height of circumfluence method and ultrasonic extraction in the extract of the present invention, and peak number is also more in the extract, illustrates that composition in the extract of the present invention is more than the extract of reflux, extract, and the extract of supersound extraction.
(2) effect for reducing blood fat of the extract of Different Extraction Method
1.1 experiment material
96 of male SD rats (quality certification SCXK (Guangdong) 2003-0002)
XUEZHIKANG JIAONANG (Beijing big dimension letter bio tech ltd, the accurate word of traditional Chinese medicines: Z10950029, lot number: 20080110);
Cholesterol reagent box (glad biotechnology research institute of Shanghai section, lot number: 20080301);
High density lipoprotein test kit (glad biotechnology research institute of Shanghai section, lot number: 20080301);
Low density lipoprotein, LDL test kit (glad biotechnology research institute of Shanghai section, lot number: 20080301);
Triglyceride test kit (glad biotechnology research institute of Shanghai section, lot number: 20080301);
High glucose and high fat feed formula: 81% normal feedstuff, 5% sodium cholate, 1.5% cholesterol, 2% yolk powder, 3% sugar, 10% Adeps Sus domestica, 25g mineral, 50g compound vitamin;
Exsiccant Semen Cassiae medical material (purchasing) in Guangzhou medical material company;
The extract of Semen Cassiae Different Extraction Method (self-control);
Instrument: visible spectrophotometer (the general all purpose instrument company limited of analysing in Beijing).
1.2 the preparation of Semen Cassiae extract
Identical with the 4th the described extracting method of (one) part.
1.3 experimental technique
96 of male SD rats, adaptability are divided into 8 groups at random by body weight after raising a week, 12 every group, are respectively: normal control group, model control group, Xuezhikang matched group 0.11g/kgd
-1, high pressure extracts high dose group 6.97g/kgd continuously
-1, high pressure extract continuously in dosage group: 2.31g/kgd
-1, high pressure extracts low dose group: 0.77g/kgd continuously
-1, supersound extraction group 2.31g/kgd
-1Reflux, extract, group 2.31g/kgd
-1Feed in formal experiment first day and to carry out modeling to the high glucose and high fat feedstuff, and inject carbon tetrachloride vegetable oil mixed liquor (60% vegetable oil for each modeling group rat muscle, 40% carbon tetrachloride) 3ml/kg, each group gives the medicine filling stomach of corresponding dosage simultaneously, the normal control group gives the equal-volume normal saline, continuous 8 weeks, water is can't help in fasting in preceding 12 hours of inferior morning, last was irritated behind the stomach 40 minutes, with 3% pentobarbital sodium 0.12ml/100g anesthesia, cut open the belly, abdominal aortic blood free liver, 10% formalin pathology to be done is made film, is fixed in normal saline 2ml flushing.(2000r/min 10min) gets serum to centrifugal blood, detects and respectively organizes rat blood serum T-CHOL (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein, LDL (LDL) level, and the animal liver index is observed the rat liver metamorphosis.
1.4 evaluation of pesticide effectiveness result relatively
1.4.1 rats'liver assessment of indices result
Table 3 rats'liver assessment of indices result
Annotate:
#P＜0.05,
##P＜0.01 is compared with normal group;
*P＜0.05,
*P＜0.01 is compared with model group;
△P＜0.05,
△ △P＜0.01 is compared with the high pressure extract high dose group;
★P＜0.05 is compared with the high pressure extract low dose group.
By table 3 and Fig. 8 as seen, model group liver index is obviously than normal group height, high pressure extracts the effect that each dosage group all has blood fat reducing continuously, and the drug effect of reflux, extract, group and supersound extraction group is obvious not as high pressure extracts each dosage group continuously, even does not see obvious reduction liver index effect.And extract continuously in each dosage group at high pressure, it is more obvious than the effect of low dose group that high dose group reduces the exponential effect of liver.
1.4.2 rat blood serum T-CHOL (TC) measurement result
Table 4 rat blood serum T-CHOL measurement result
Annotate:
#P＜0.05,
##P＜0.01 is compared with normal group;
*P＜0.05,
*P＜0.01 is compared with model group
By table 4 and Fig. 9 as seen, each administration group all has apparent in view effect to reducing the rat blood serum cholesterol, and wherein the effect of reflux, extract, group is best, and high pressure extracts high dose group continuously and low dose group is taken second place.
1.4.3 rat high density lipoprotein (HDL) level determination result
Table 5 rat high density lipoprotein (HDL) level determination result
Annotate:
#P＜0.05,
##P＜0.01 is compared with normal group;
*P＜0.05,
*P＜0.01 is compared with model group;
△P＜0.05 is compared with dosage group in the high pressure extract;
★P＜0.05,
★ ★P＜0.01 is compared with the high pressure extract low dose group.
From the result of table 5 and Figure 10 reflection, each administration group all has the effect of high density lipoprotein increasing (HDL) level.The effect of supersound extraction group, reflux, extract, group and Xuezhikang matched group is suitable.High pressure extracts each dosage group continuously all has obvious effects to rising rat high density lipoprotein (HDL) level, and its effect size is followed successively by: high pressure extract continuously high dose group＜high pressure extract continuously in dosage group＜high pressure extract low dose group continuously.
1.4.4 rat low density lipoprotein, LDL (LDL) level determination result
Table 6 rat low density lipoprotein, LDL (LDL) level determination result
Annotate:
#P＜0.05,
##P＜0.01 is compared with normal group;
*P＜0.05,
*P＜0.01 is compared with model group
By table 6 and Figure 11 as seen, each administration group all has positive effect to reducing rat low density lipoprotein, LDL (LDL).Wherein, reflux, extract, group, high pressure extract continuously high dose group, high pressure extract continuously in dosage group and high pressure to extract the effect of low dose group continuously all obvious than the Xuezhikang matched group; And the dosage group is suitable with reflux, extract, group effect in the continuous extraction of high pressure; The effect that high pressure extracts high dose group continuously is the most remarkable, and the level of rat low density lipoprotein, LDL (LDL) is reduced to normal level.
1.4.5 rat blood serum triglyceride (TG) assay result
Table 7 rat blood serum triglyceride (TG) assay result
Annotate:
#P＜0.05,
##P＜0.01 is compared with normal group;
*P＜0.05,
*P＜0.01 and model group
By table 7 and Figure 12 as seen, each administration group all has the effect that reduces rat blood serum triglyceride (TG) content, and effect is suitable.
3.3 conclusion
By table 3, table 4, table 5, table 6, table 7 and Fig. 8 thereof～12 as seen, the equal highly significant of TC, TG, LDL-C of hyperlipidemia model group mice serum is higher than normal control group (P＜0.01), and two groups HDL difference with insignificance (P＞0.05).The content and the utmost point that positive Xuezhikang matched group can reduce TC (P＜0.05) significantly reduce the content of LDL-C (P＜0.01) significantly.This shows that the hyperlipemia in mice model that this experiment is set up is successful.
Each administration group is all influential to TC, TG and the LDL of mice serum, can make it that to a certain degree decline is arranged, but have only high pressure to extract each dosage group continuously HDL is had the rising effect.Except to reducing aspect serum triglycerides (TG) content, the effect that reflux, extract, group, supersound extraction group and high pressure extract each dosage group continuously quite outside, it is the most remarkable that the level of the TC that reduces mice serum and LDL and rising HDL level are all extracted each dosage group effect continuously with high pressure.
Cumulated volume experiment and experimental result, the Semen Cassiae extract that high pressure extracts continuously is the drug effect of blood fat reducing obviously, and the effect of treatment hyperlipemia obviously is better than compound preparation-Xuezhikang.
From above result, can draw and the invention has the advantages that:
(1) as seen from the above-mentioned test of pesticide effectiveness, Semen Cassiae extract of the present invention has the drug effect of the blood fat reducing stronger than the extract of other extracting method extractions, and its effect in the treatment hyperlipemia obviously is better than compound preparation-Xuezhikang, therefore can not need and the other drug compatibility, and be prepared into the Semen Cassiae single preparations of ephedrine that is used for blood fat reducing, and has the effect of tangible blood fat reducing.
(2) composition (referring to Fig. 1～3 and table 1) that contains in the Semen Cassiae extract of each constituents that contains in the Semen Cassiae extract of the present invention more than traditional reflux extraction and ultrasonic extraction gained.And in Semen Cassiae extract of the present invention, isolate the new main component of 5 kinds of blood fat reducing: 1,2,6,8-tetrahydroxy-7-methoxyl group-3-tectoquinone (molecular weight 316); 1,2,6-trihydroxy-7,8-dimethoxy-3-tectoquinone (molecular weight 330); 2-hydroxyl-1,6,7,8-tetramethoxy-3-tectoquinone (molecular weight 358); 2,6-dihydroxy-1,7,8-trimethoxy-3-tectoquinone (molecular weight 344); 1,2-dihydroxy-6,7,8-trimethoxy-3-tectoquinone (molecular weight 344).
(3) Semen Cassiae extract of the present invention is to adopt the high pressure continuous extraction method to extract gained, and high pressure extract device and external environment have good heat exchange, in whole leaching process, can keep at room temperature and carry out, thereby farthest kept the natural structure of pharmacological active substance and various nutritional labelings in the Chinese crude drug, problem such as the effective ingredient degeneration of having avoided causing, loss, pharmacologically active reduction because of heat effect.
Description of drawings
Fig. 1 is the high pressure HPLC chromatogram of the Semen Cassiae extract of extraction continuously;
Fig. 2 is the total ions chromatogram of Semen Cassiae extract of the present invention;
Fig. 3 is the HSCCC separate colors spectrogram of the n-butanol extract of Semen Cassiae extract of the present invention;
Fig. 4-the 1st, the high-efficient liquid phase chromatogram of peak I gleanings in the HSCCC separate colors spectrogram of the n-butanol extract of Semen Cassiae extract of the present invention;
Fig. 4-the 2nd, the high-efficient liquid phase chromatogram of peak II gleanings in the HSCCC separate colors spectrogram of the n-butanol extract of Semen Cassiae extract of the present invention;
Fig. 4-the 3rd, the high-efficient liquid phase chromatogram of peak III gleanings in the HSCCC separate colors spectrogram of the n-butanol extract of Semen Cassiae extract of the present invention;
Fig. 4-the 4th, the high-efficient liquid phase chromatogram of peak IV gleanings in the HSCCC separate colors spectrogram of the n-butanol extract of Semen Cassiae extract of the present invention;
Fig. 4-the 5th, the high-efficient liquid phase chromatogram of peak V gleanings in the HSCCC separate colors spectrogram of the n-butanol extract of Semen Cassiae extract of the present invention;
Fig. 5 is the HPLC finger printing of the Semen Cassiae extract of reflux, extract, method extraction;
Fig. 6 is the HPLC finger printing of the Semen Cassiae extract of ultrasonic extracting method extraction;
Fig. 7 is the high pressure HPLC finger printing of the Semen Cassiae extract of extraction continuously;
Fig. 8 is the exponential bar diagram as a result of the rats'liver of each test group;
Fig. 9 is the bar diagram as a result of the rat blood serum T-CHOL of each test group;
Figure 10 is the bar diagram as a result of the rat hdl level of each test group;
Figure 11 is the bar diagram as a result of the rat low-density lipoprotein white level of each test group;
Figure 12 is the bar diagram as a result of the rat blood serum triglyceride of each test group;
Figure 13 is the partial structurtes sketch map of high pressure extract device.
The specific embodiment
Following embodiment just is used for content of the present invention is set forth, rather than restriction, therefore in implication suitable with claims of the present invention and any change in the scope, all should think to be included in the scope of claims.
The structure of high pressure extract device (Figure 13): described high pressure extract device comprises the high-pressure pump (not shown), be provided with the valve body 84 of cavity, be provided with the valve seat 81 of through hole, collision ring 82 and the spool 83 that is connected with the high pressure spring, spool 83 is positioned at an end of valve cavity, the other end of valve cavity is equipped with valve seat 81, have liquid-inlet 811 and liquid outlet 813 on the valve body 84, the high pressure delivery side of pump is communicated with the liquid-inlet 811 of valve body 84,811 of the liquid-inlets of valve body 84 are communicated with through hole in the valve seat 81, valve seat 81 is a boss structure, it has shoulder, collision ring 82 and valve seat 81 coaxial being fixed in the valve body 84, collision ring 82 is positioned at the end of valve seat shoulder, valve seat 81 in valve cavity end face and spool end between leave the flow gap that flows through for liquid, this gap is positioned at the inwall middle part of collision ring 82, part collision ring is nested with spool, make the annular working gap 812 that leaves the collision feed liquid between the two, liquid outlet 813 on the described valve body 84 is connected with annular working gap 812, be used to discharge the feed liquid after high pressure breaks, this annular working gap 812 is in the scope of 0.1～1mm.
Embodiment one:
Depend on that pine torch powder 100g mixes immersion 60 minutes with 1000ml70% ethanol; Then the feed liquid of gained is sent in the high pressure extract device, feed liquid is passed through the annular working gap of high pressure extract device continuously under the effect of 90Mpa high pressure, on the collision ring of high speed impact in the high pressure extract device, make the cell wall rupture of Semen Cassiae, effective ingredient is spread in the solvent; Take out the feed liquid through HIGH PRESSURE TREATMENT at last, filter, remove the material slag, filtrate concentrates through reclaiming ethanol, and drying obtains the extract powder of Semen Cassiae extract.Depend on pine torch extract powder 20g then, make tablet by known tabletting technology and equipment.
Embodiment two:
Depend on that pine torch powder 200g mixes immersion 30 minutes with 6000ml70% ethanol; Then the feed liquid of gained is sent in the high pressure extract device, feed liquid is passed through the annular working gap of high pressure extract device continuously under the effect of 60Mpa high pressure, on the collision ring of high speed impact in the high pressure extract device, make the cell wall rupture of Semen Cassiae, effective ingredient is spread in the solvent; Take out the feed liquid through HIGH PRESSURE TREATMENT at last, filter, remove the material slag, filtrate concentrates the extractum that obtains Semen Cassiae extract through reclaiming ethanol.Depend on pine torch extractum 35g then, make granule by the technology and equipment of known system granule.
Embodiment three:
Depend on that pine torch powder 500g mixes immersion 60 minutes with 6000ml60% ethanol; Then the feed liquid of gained is sent in the high pressure extract device, feed liquid circulates continuously under the effect of 50Mpa high pressure and repeatedly passes through the annular working gap of high pressure extract device, so that the cell wall that makes Semen Cassiae is through multiple impact, the circulation time is 15min continuously, make the cell wall rupture of Semen Cassiae, effective ingredient is spread in the solvent; Take out the feed liquid through HIGH PRESSURE TREATMENT at last, filter, remove the material slag, filtrate concentrates through reclaiming ethanol, and drying obtains the extract powder of Semen Cassiae extract.Depend on pine torch extract powder 90g then, make tablet by known tabletting technology and equipment.
Embodiment four:
Depend on that pine torch powder 500g mixes immersion 50 minutes with 7500ml80% ethanol; Then the feed liquid of gained is sent in the high pressure extract device, feed liquid circulates continuously under the effect of 80Mpa high pressure and repeatedly passes through the annular working gap of high pressure extract device, so that the cell wall that makes Semen Cassiae is through multiple impact, the circulation time is 30min continuously, make the cell wall rupture of Semen Cassiae, effective ingredient is spread in the solvent; Take out the feed liquid through HIGH PRESSURE TREATMENT at last, filter, remove the material slag, filtrate concentrates through reclaiming ethanol, and drying obtains the extract powder of Semen Cassiae extract.Depend on pine torch extract powder 82g then, make granule by the technology and equipment of known preparation granule.
Embodiment five:
Depend on that pine torch powder 500g mixes immersion 50 minutes with 6000ml50% ethanol; Then the feed liquid of gained is sent in the high pressure extract device, feed liquid circulates continuously under the effect of 70Mpa high pressure and repeatedly passes through the annular working gap of high pressure extract device, so that the cell wall that makes Semen Cassiae is through multiple impact, the circulation time is 20min continuously, make the cell wall rupture of Semen Cassiae, effective ingredient is spread in the solvent; Take out the feed liquid through HIGH PRESSURE TREATMENT at last, filter, remove the material slag, filtrate concentrates through reclaiming ethanol, and drying obtains the extract powder of Semen Cassiae extract.Depend on pine torch extract powder 80g then, make capsule by the technology and equipment of known preparation capsule.
Embodiment six
Depend on that pine torch powder 200g mixes immersion 30 minutes with 6000ml70% ethanol; Then the feed liquid of gained is sent in the high pressure extract device, feed liquid circulates continuously under the effect of 60Mpa high pressure and repeatedly passes through the annular working gap of high pressure extract device, so that the cell wall that makes Semen Cassiae is through multiple impact, the circulation time is 30min continuously, make the cell wall rupture of Semen Cassiae, effective ingredient is spread in the solvent; Take out the feed liquid through HIGH PRESSURE TREATMENT at last, filter, remove the material slag, filtrate concentrates through reclaiming ethanol, and drying obtains the extract powder of Semen Cassiae extract.Depend on pine torch extract powder 40g then, make capsule by the technology and equipment of known preparation capsule.
Embodiment seven
Depend on that pine torch powder 300g mixes immersion 40 minutes with 7500ml60% ethanol; Then the feed liquid of gained is sent in the high pressure extract device, feed liquid circulates continuously under the effect of 30Mpa high pressure and repeatedly passes through the annular working gap of high pressure extract device, so that the cell wall that makes Semen Cassiae is through multiple impact, the circulation time is 25min continuously, make the cell wall rupture of Semen Cassiae, effective ingredient is spread in the solvent; Take out the feed liquid through HIGH PRESSURE TREATMENT at last, filter, remove the material slag, filtrate concentrates the extractum that obtains Semen Cassiae extract through reclaiming ethanol.Depend on pine torch extractum 50g then, make granule by the technology and equipment of known preparation granule.
Embodiment eight
Depend on that pine torch powder 300g mixes immersion 30 minutes with 6000ml85% ethanol; Then the feed liquid of gained is sent in the high pressure extract device, feed liquid circulates continuously under the effect of 40Mpa high pressure and repeatedly passes through the annular working gap of high pressure extract device, so that the cell wall that makes Semen Cassiae is through multiple impact, the circulation time is 10min continuously, make the cell wall rupture of Semen Cassiae, effective ingredient is spread in the solvent; Take out the feed liquid through HIGH PRESSURE TREATMENT at last, filter, remove the material slag, filtrate concentrates through reclaiming ethanol, and drying obtains the extract powder of Semen Cassiae extract.Depend on pine torch extract powder 40g then, make tablet by the technology and equipment of known preparation tablet.
Claims (8)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN2009100396313A CN101549008B (en) | 2009-05-21 | 2009-05-21 | Application of cassia seed extract extracted continuously under high pressure in the preparation of blood lipid-lowering drugs |
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| CN2009100396313A CN101549008B (en) | 2009-05-21 | 2009-05-21 | Application of cassia seed extract extracted continuously under high pressure in the preparation of blood lipid-lowering drugs |
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| KR101988757B1 (en) * | 2017-12-19 | 2019-06-12 | 가톨릭대학교 산학협력단 | Composition comprising 1,2-dihydroxy-3-methylanthraquinone as an effective ingredient for preventing or treating of cancer |
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